METHODS FOR ISOLATING AND CULTURING LIVING CELLS USING METHOD OF PERMEABILIZING CELL MEMBRANE
First Claim
1. A method of isolating target cells in a living state, comprising:
- (s1) treating a test material including target cells with a streptococcal hemolytic exotoxin, i.e., Streptolysin O (SLO);
(s2) treating the test material with an antibody against a target intracellular protein of the target cells; and
(s3) isolating the target cells in a living state, which have a positive response to the antibody.
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Accused Products
Abstract
Provided are methods of isolating and culturing various cells in a living state including peripheral blood mononuclear cells (PBMCs) isolated from peripheral blood, which use a cell membrane permeabilization method. The methods use a streptococcal hemolytic exotoxin, which binds to cholesterol present in a cell membrane so as to make a pore therein, thereby allowing an exogenous protein to be permeated into the cell, and is a technique of isolating and culturing desired cells in a living state by probing a specific intracellular protein and performing flow cytometry (FACS). According to the methods, various intracellular proteins that could not previously be isolated and cultured from a patient'"'"'s blood as well as various tissues may be targeted, and homogeneous cells may be obtained with high purity and high efficiency. Therefore, it is expected that the methods will highly contribute to many applications targeting a specific intracellular protein and research on mechanisms of various diseases and treatment thereof.
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Citations
13 Claims
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1. A method of isolating target cells in a living state, comprising:
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(s1) treating a test material including target cells with a streptococcal hemolytic exotoxin, i.e., Streptolysin O (SLO); (s2) treating the test material with an antibody against a target intracellular protein of the target cells; and (s3) isolating the target cells in a living state, which have a positive response to the antibody. - View Dependent Claims (2, 3, 4, 5, 6, 12)
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7. A method of isolating target cells in a living state, comprising:
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(s1) treating a test material including target cells with a streptococcal hemolytic exotoxin, i.e., Streptolysin O (SLO); (s2) treating the test material with an antibody against a target intracellular protein of the target cells; and (s3) isolating the target cells in a living state, which have a positive response to the antibody through fluorescence-activated cell sorting (FACS) analysis. - View Dependent Claims (8, 9, 10, 11, 13)
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Specification